Double-layer composite support for establishing full-thickness skin and preparation method of double-layer composite support

A double-layer compound, full-thickness skin technology, applied in medical science, prostheses, etc., can solve the problems of long culture period of full-thickness skin model, poor mechanical properties of collagen gel scaffold, easy enzymatic hydrolysis, etc., and achieve tissue culture cycle Short, low cost, good biocompatibility effect

Inactive Publication Date: 2018-08-10
GUANGDONG UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0007] In view of this, a double-layer composite scaffold for constructing full-thickness skin and its preparation method of the present invention can effectively solve the poor mechanical properties of the current collagen gel scaffold, and the collagen gel scaffold is easy to be enzymatically degraded during the tissue culture process. , and the technical defect of a long culture period for constructing a full-thickness skin model with a porous sponge structure scaffold

Method used

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  • Double-layer composite support for establishing full-thickness skin and preparation method of double-layer composite support
  • Double-layer composite support for establishing full-thickness skin and preparation method of double-layer composite support
  • Double-layer composite support for establishing full-thickness skin and preparation method of double-layer composite support

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preparation example Construction

[0048] This specific embodiment discloses a method for preparing a double-layer composite scaffold for constructing full-thickness skin, including:

[0049] Step 1, mixing degradable protein with citric acid solution and freezing at low temperature, followed by vacuum freeze-drying to obtain a porous sponge precursor, and cross-linking the porous sponge precursor at high temperature to obtain a porous sponge scaffold;

[0050] Step 2, electrospinning the biodegradable fiber on the surface of the porous sponge support to obtain a nanofiber layer;

[0051] Step 3: After cross-linking the porous sponge scaffold with the nanofiber layer attached to the surface, a double-layer composite scaffold for constructing full-thickness skin is prepared.

[0052] Wherein, the degradable protein includes one or more of gelatin, collagen, silk protein, elastin and fibrin.

[0053] Wherein, the biodegradable fiber includes one or more of gelatin, collagen, silk protein, polylactic acid, polyam...

Embodiment 1

[0061] The preparation method of the double-layer composite scaffold for constructing full-thickness skin according to the embodiment of the present invention is as follows:

[0062] (1) Mix liquid polydimethylsiloxane (PDMS) and curing agent in a mass ratio of 5:1, stir and mix well, weigh 10g and completely cover the bottom area of ​​125cm 2 In a circular dish, vacuum out the air bubbles and put it in an oven at 80°C to cure polydimethylsiloxane (PDMS), then punch holes with a circular punch with a diameter of 12cm, and then punch holes with a circular hole with a diameter of 8cm The ring-shaped PDMS is obtained by punching holes, and finally the surface is modified with a plasma surface processor to form a mold.

[0063] (2) Dissolve a certain amount of gelatin and citric acid in deionized water, wherein the solid content of gelatin is 1%, and the solid content of citric acid is 0.1%. Stir evenly with a magnetic stirrer at room temperature to obtain a gelatin solution , ad...

Embodiment 2

[0071] This example uses the double-layer composite scaffold for constructing full-thickness skin prepared in Example 1, the gelatin porous scaffold of the prior art and human keratinocytes to construct full-thickness skin. Figure 4 and Figure 5For the experimental results, Figure 4 Human keratinocytes were simultaneously seeded on the porous sponge scaffold (S1) and the nanofibrous layer (S2) of the two-layer composite scaffold for constructing full-thickness skin one day and three days later Electron microscope images, as can be seen from the figure, Figure 4 In (a) and (b), the mass is the form of keratinocytes adhering and proliferating on the S1 porous sponge scaffold; (c) and (d), the black sheet-like part is the form of keratinocytes on the S2 nanofiber layer, wherein The raised single globular structure is the nucleus. Inoculation of human keratinocytes in S2 compared with S1, the cells on the surface of the S2 nanofiber layer adhered better and the nanofibers p...

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Abstract

The invention belongs to the technical field of tissue engineering, and particularly relates to a double-layer composite support for establishing a full-thickness skin and a preparation method of thedouble-layer composite support. The preparation method comprises the following steps: I, mixing degradable proteins and a citric acid solution, freezing at a low temperature, and vacuum freeze dryingto obtain a porous sponge precursor, and cross-linking the porous sponge precursor at a high temperature to obtain a porous sponge support; II, mixing biodegradable fibers and a first solvent to obtain a biodegradable fiber solution, mixing the biodegradable fiber solution and a second solvent, and performing electrostatic spinning on the surface of the porous sponge support to obtain a nano fiberlayer; and III, enabling the porous sponge support with the nano fiber layer adhered on the surface to be cross-linked, and thus obtaining the double-layer composite support for establishing the full-thickness skin. By adopting the double-layer composite support, the technical defects that a collagen gel support is poor in mechanical performance, the collagen gel support is easy to perform enzymolysis in the tissue culture process, and the culture period of the collagen gel support for establishing a full-thickness skin model is long can be solved.

Description

technical field [0001] The invention belongs to the technical field of tissue engineering, and in particular relates to a double-layer composite scaffold for constructing full-thickness skin and a preparation method thereof. Background technique [0002] The skin is the largest organ tissue in the human body. It can block the invasion of foreign substances, protect moisture, and regulate and sense temperature. Human skin is mainly composed of epidermis and dermis, wherein the epidermis has multiple layers of dense cells to play a barrier function, and the dermis is mainly composed of fibroblasts, collagen fibers and elastic fibers, which have certain mechanical strength and elasticity. [0003] Bacteria and pathogens are easy to invade the skin and cause infection, which can lead to death in severe cases. In the past 30 years, a lot of research has been carried out on the repair of skin damage, especially the research on scaffolds, and a variety of products have been launch...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61L27/40A61L27/60A61L27/22A61L27/18A61L27/50A61L27/56A61L27/58
CPCA61L27/18A61L27/222A61L27/225A61L27/227A61L27/50A61L27/56A61L27/58A61L27/60A61L2300/412C08L89/00C08L67/04C08L77/04
Inventor 杜志云黄博鑫汤亚东董长治郑雅婷
Owner GUANGDONG UNIV OF TECH
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